| Objective:The aim of this study was to investigate the molecular characterization and gene mutation spectrum of thalassemia in Fuzhou area to provide information for comprehensive thalassemia screening,prenatal diagnosis and genetic counselling.Methods:The study was retrospective and cross-sectional involving 1369 patients.All the patients were clinically suspected for thalassemia,or showed abnormal results in red blood cell indices analysis or hemoglobin electrophoresis analysis.The cohort of patients included 681pregnant women,who were selected from 7946 pregnant women according to the screening indices of MCV and MCH?MCV<80fl and MCH<27pg?.Genomic DNA was extracted from peripheral blood samples,and analyzed for thalassemia mutations by traditional methods of Gap-PCR and PCR-Reverse Dot blot assays.Next generation sequencing?NGS?was further applied to detect thalassemia mutations which were undetectable by the traditional methods.The mutations detected by NGS were validated with Sanger sequencing or real-time PCR or another Gap-PCR methods.Results:gene mutations of 743 thalassemia were identified,in which 733 cases were detected by traditional methods of Gap-PCR and PCR-Reverse Dot blot assay,and the rest of the 10 by NGS.4.43%of pregnant women?352/7946?were found with a gene mutation of thalassemia.The total of 743 mutations were detected including 422 cases with?,292 cases with?,and 9 cases with?/?thalassemia gene mutations respectively.Among these,--SEA/??and IVS-II-654?C>T?were the most frequent mutations.Four rare mutations were identified by NGS;including one codon14/15?+G??thalassemia mutation,2 cases of?globin gene duplication mutation???/??and one1-HBB?gene deletion mutation.Conclusion:1.The prevalence of thalassemia in Fuzhou area was 4.43%in the present study.2.The mutation genotypes of thalassemia in Fuzhou area were many and various.3.Undetectable common and rare gene mutations of thalassemia by traditional methods could be revealed by NGS. |
PDF Full Text Request